经膈肌穿刺法快速准确评估大鼠肺动脉高压

Rapid and accurate assessment of pulmonary hypertension in rats via diaphragmatic puncture method

目的构建一种操作简单快捷、测量准确的大鼠右心室压力测定方法。方法采用随机方法将30只8~10周龄的雄性SD大鼠分为3组:经胸右心室穿刺测压组(trans-thoracic right ventricular puncture manometry group,TTM组,n=10),经膈肌穿刺测压组(trans-diaphragmatic puncture manometry group,TDM组,n=10)和野百合碱诱导肺动脉高压采用经膈肌穿刺测压组(monocrotaline-induced pulmonary hypertension with trans-diaphragmatic manometry group,MCT-TDM组,n=10)。MCT-TDM组经胸腹侧皮下注射野百合碱(60 mg/kg),TTM组和TDM组注射等体积的无菌生理盐水。模型建立4周后,使用经膈肌穿刺法测定TDM组和MCT-TDM组大鼠的右心室收缩压(right ventricular systolic pressure,RVSP),随后立即开胸验证穿刺点的准确性;使用经胸右心室穿刺法测定TTM组大鼠RVSP。比较两种测压方法操作时间、测压成功率、RVSP等指标。取大鼠心肺组织,分离左、右心室,并计算右心室肥厚指数(right ventricular hypertrophy index,RVHI)。最后,对左肺组织进行苏木精-伊红(hematoxylin eosin,HE)染色,评价肺部小血管病变程度。结果TTM组从麻醉开始到成功测出右心室压力的时间为(6.83±1.36)min,TDM组为(8.13±1.25)min,MCT-TDM组为(8.32±1.23)min。TTM组测压成功率为70%,TDM组测压成功率为100%,MCT-TDM组成功率为90%。TDM组和TTM组测得的RVSP、RVHI差异无统计学意义(P>0.05)。MCT-TDM组的RVSP及RVHI均显著高于TDM组(P<0.001)。MCT-TDM组大鼠肺小动脉管壁相较于TDM组明显增厚,管腔狭窄。结论采用膈肌穿刺法测量肺动脉高压大鼠的右心室压力,这种方法无需开胸,避免了昂贵设备的使用,能准确、快速、简便地测量右心室血流动力学,具有较高的推广价值。

Objective To establish a simple,rapid and accurate method to measure the right ventricular pressure in rats.Methods A total of 30 male SD rats aged 8-10 weeks were randomly divided into 3 groups:the trans-thoracic right ventricular puncture manometry group(TTM group,n=10),trans-diaphragmatic puncture manometry group(TDM group,n=10),and the monocrotaline-induced pulmonary hypertension with trans-diaphragmatic manometry group(MCT-TDM group,n=10).The MCT-TDM group underwent subcutaneous injection of monocrotaline through the thoracoabdominal side(60 mg/kg),while the TTM and TDM groups were injected with an equivalent volume of sterile saline.Four weeks after model establishment,the right ventricular systolic pressure(RVSP)of the TDM and MCT-TDM groups was measured using the trans-diaphragmatic puncture method,followed by thoracotomy to verify the accuracy of the puncture site;RVSP of the TTM group was measured using the trans-thoracic right ventricular puncture method.The operation time,success rate,and RVSP were compared between the two manometry methods.The hearts and lung tissues of the rats were extracted,left and right ventricles were separated,and the right ventricular hypertrophy index(RVHI)was calculated.Lastly,the left lung tissue was stained with hematoxylin and eosin(HE)to evaluate the extent of pulmonary microvascular lesions.Results The time taken from anesthesia to successful measurement of RVSP was(6.83±1.36)minutes for the TTM group,(8.13±1.25)minutes for the TDM group,and(8.32±1.23)minutes for the MCT-TDM group.The success rate were 70%for the TTM group,100%for the TDM group,and 90%for the MCT-TDM group.There were no significant differences in RVSP and RVHI between the TDM and TTM groups(P>0.05).The RVSP and RVHI in the MCT-TDM group were significantly higher than those in the TDM group(P<0.001).In the MCT-TDM group,the pulmonary arterioles showed notable thickening of the vessel walls and narrowing of the lumens.Conclusion Measuring the right ventricular pressure with diaphragmatic puncture technique negates the need for thoracotomy and circumvents the use of costly equipment.It enables precise,swift,and straightforward assessment of the right ventricular hemodynamics,holding considerable value for widespread application.